Abnormal hemoglobins come to the attention of this laboratory by virtue of its serving as a reference laboratory for the Michigan region. The structural abnormalities of abnormal hemoglobins are defined and correlated with function and kinetics of synthesis of the molecule. From efforts such as these this laboratory noted a trimodal distribution in the proportion of HbG-alpha Philadelphia in heterozygotes with models at 20, 30, and 40% HbG. Synthesis of alpha and beta chains is balanced. The hypothesis will be tested that this distribution reflects varying numbers of alpha chain structured loci exclusive of alpha thalassemia. The alpha/beta ratio of cDNA-DNA nucleic acid hybridizations will be measured for the three phenotypes to measure gene dosage directly. Hb Wayne, an alpha chain frameshift mutant, will also be studied. This mutant will also be studied. This mutant is a minor component with balanced synthesis (alpha A - alpha W /beta equals 1). The mechanism of balance both of these will be explored by translating mRNA in the wheat germ system to determine whether transcription is balanced. In addition the alpha/beta ratio by cDNA-RNA will be measured in nucleic and cytoplasm to evaluate mRNA processing. Abnormal hemoglobins will be examined by a new heat denaturation method to determine which are superstable. In addition fetal hemoglobin and carbonic anhydrase will be measured by a cytoimmunofluorescence technique to determine whether the inverse correlation between the two noted in sickle cell anemia whole blood is present in individual cells. Attempts will be made to stimulate fetal hemoglobin synthesis in cultured human bone marrow cells and relate it to carbonic anhydrase synthesis. Attempts will also be made to stimulate fetal hemoglobin synthesis in hamsters, both in vivo and in cultured marrow cells.